Recombinant Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein (S) (N354D, D364Y) is produced in a mammalian cell expression system, ensuring proper protein folding and post-translational modifications. This product features a partial protein length from amino acids 319 to 541 with specific mutations at positions N354D and D364Y, and is C-terminally tagged with 10xHis for ease of purification and detection. The protein maintains a purity level greater than 90% as verified by SDS-PAGE.
The Spike glycoprotein of SARS-CoV-2 plays a crucial role in the virus's ability to infect host cells. It mediates viral entry by binding to the host cell receptor, angiotensin-converting enzyme 2 (ACE2). Given its essential function in viral pathogenesis, the Spike protein has become a key target for vaccine development and therapeutic interventions. This makes it a significant focus of research in understanding and combating COVID-19.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. Receptor Binding Domain (RBD) Interaction Studies
This recombinant SARS-CoV-2 spike protein fragment covers the receptor binding domain (amino acids 319-541) and can be used to study interactions with the human ACE2 receptor in vitro. The N354D and D364Y mutations may alter binding affinity or specificity compared to the wild-type protein, which makes this variant valuable for comparative binding studies. The C-terminal His-tag allows for purification and immobilization for surface plasmon resonance, bio-layer interferometry, or ELISA-based binding assays. Researchers can quantify binding kinetics, determine dissociation constants, and assess how these specific mutations impact receptor recognition.
2. Neutralizing Antibody Development and Screening
The RBD region appears to be a critical target for neutralizing antibodies. This mutant variant can serve as an antigen for antibody discovery and characterization studies. The His-tag makes protein capture straightforward in screening assays such as phage display, hybridoma screening, or single-cell antibody sequencing platforms. Researchers can evaluate whether existing neutralizing antibodies maintain efficacy against these mutations or identify novel antibodies specific to this variant. This application is particularly relevant for understanding immune escape mechanisms and developing broadly neutralizing antibody candidates.
3. Structural and Biophysical Characterization
The defined amino acid region (319-541) with specific mutations provides a well-characterized protein fragment for structural studies. X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy could be used to examine this protein. The mammalian expression system ensures proper glycosylation and folding patterns that may be critical for maintaining native-like structure. Researchers can investigate how the N354D and D364Y mutations affect protein conformation, stability, and dynamics compared to wild-type RBD. The His-tag can be used for protein purification and concentration determination in these biophysical analyses.
4. Protein-Protein Interaction Mapping
The His-tagged RBD variant can be used in pull-down assays to identify and characterize cellular proteins that interact with the SARS-CoV-2 spike protein beyond ACE2. By using the His-tag for immobilization on nickel-affinity matrices, researchers can incubate the protein with cell lysates or purified protein libraries to discover novel binding partners. The specific mutations may reveal altered interaction profiles that could provide insights into viral pathogenesis mechanisms. Mass spectrometry analysis of pulled-down complexes can identify both known and previously uncharacterized spike-interacting proteins.
5. Immunogenicity and Vaccine Research Applications
This mutant RBD fragment appears suitable as a research tool for evaluating immune responses in preclinical vaccine studies. The protein can be used to immunize laboratory animals to assess the immunogenicity of these specific mutations and compare antibody responses to wild-type versus mutant antigens. The His-tag allows for consistent protein quantification and quality control across immunization studies. Researchers can analyze whether vaccines based on earlier SARS-CoV-2 variants generate cross-reactive immune responses against this mutated RBD sequence through in vitro immunoassays.
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